Influencing Factors for Cure of Clonorchiasis by Praziquantel Therapy: Infection Burden and CYP3A5 Gene Polymorphism

Chemotherapy of clonorchiasis with praziquantel (PZQ) is effective but about 15% of treated cases have been reported uncured. The present study investigated correlation of single nucleotide polymorphisms (SNPs) of the cytochrome P450 gene, CYP3A5 and cure of clonorchiasis. A total of 346 egg passing residents were subjected and treated by 3 doses of 25 mg/kg PZQ. Reexamination recognized 33 (9.5%) uncured and 313 cured. Numbers of eggs per gram of feces (EPGs) before treatment were significantly lower in the cured group than in the uncured group (2,011.2+/-3,600.0 vs 4,998.5+/-7,012.0, PA and g.27526C>T of CYP3A5 were 15.2% and 9.1% while those were 3.8% and 1.0%, respectively, in the cured group. The cure rate was significantly lower in the cases with SNP at g.27526C>T and EPGs> or =1,000. In conclusion, EPGs and SNPs of CYP3A5 are factors which influence cure of clonorchiasis by PZQ therapy. It is strongly suggested to recommend 2-day medication for individuals with high EPGs> or =1,000.

Collection of Clonorchis sinensis adult worms from infected humans after praziquantel treatment

A cohort was established for evaluation of cancer risk factors in Sancheong-gun, Gyeongsangnam-do, Korea. As one of the cohort studies, stools of 947 residents (403 males and 544 females, age range: 29-86 years) were screened for Clonorchis sinensis eggs using both Kato-Katz method and formalin-ether sedimentation technique. The overall egg positive rate of C. sinensis was 37.7% and individual EPG (eggs per gram of feces) counts ranged from 24 to 28,800. Eight egg positive residents voluntarily joined a process of collection of the passed worms after praziquantel treatment. A total of 158 worms were recovered from 5 of the 8 treated persons, ranged from 3 to 108 in each individual. The worms were 15-20 mm x 2-3 mm in size, and showed brown-pigmented, red, or white body colors. This is the first collection record of C. sinensis adult worms from humans through anthelmintic treatment and purgation. The adult worms of C. sinensis may be paralyzed by praziquantel and then discharged passively through bile flow in the bile duct and by peristaltic movement of the bowel.

Susceptibility of experimental animals to reinfection with Clonorchis sinensis

The present study observed the resistance to reinfection with Clonorchis sinensis in various experimental animals including mice, guinea pigs, rabbits, and dogs, as well as rats and hamsters. The resistance rates to reinfection in rats, mice, hamsters, guinea pigs, rabbits, and dogs were 79.7%, 58.0%, -12.6%, 54.8%, 62.6%, and 6.0%, respectively. Worms recovered from reinfected rats and mice were immature, and significantly smaller than those from the primarily infected (P < 0.01), whereas those from other animals were fully matured to adults. These findings indicate that the protective response against reinfection with C. sinensis is prominent in rats and mice, and that they may be a good animal model to investigate the mechanism of resistance to reinfection with C. sinensis.

Therapeutic evaluation of sustained-releasing praziquantel (SRP) for clonorchiasis: Phase 1 and 2 clinical studies

Sustained-releasing praziquantel (SRP) tablet was designed for single dose treatment regimen of clonorchiasis. A previous pre-clinical study confirmed its sustained-releasing characteristics and a better cure rate than conventional praziquantel (PZQ). In this clinical study, the pharmacokinetics of this SRP tablet were investigated in human volunteers (phase 1; 12 volunteers), and its curative efficacy was examined in clonorchiasis patients (phase 2; 20 volunteers). In the phase 1 clinical study, blood concentrations of both tablets showed wide individual variation. The AUC(last) of SRP was 497.9+/-519.0 ng.hr/ml (mean+/-SD) and PZQ of 628.6+/-695.5 ng.hr/ml, and the AUC(inf) of SRP was 776.0+/-538.5 ng.hr/ml and of PZQ 658.6+/-709.9 ng.hr/ml. C(max) values of SRP and PZQ were 90.7+/-82.2 ng/ml and 214.9+/-251.9 ng/ml, and T(max) values were 3.42+/-1.43 hr and 1.96+/-1.23 hr, respectively. SRP tablets showed similar AUC values, but lower C(max) and longer T(max) values than PZQ. In the phase 2 study, SRP at 30 mg/kg (single dose) achieved a 60% cure rate and a 95.5% egg reduction rate. The cure rate of a single dose SRP was unsatisfactory compared with that of the conventional PZQ dose, but much better than that achieved by a single dose PZQ.

Social and behavioral factors associated with Clonorchis infection in one commune located in the Red River Delta of Vietnam.

Social behavioral factors associated with Clonorchis infection are needed for control measures. The population in Nga Tan commune were randomly sampled and questioned to determine knowledge, perception, and health behavioral factors associated with Clonorchis infection among heads of households. The cellophane thick smear method was applied to examine their stool samples. Seven hundred and seventy-one cases were examined, the positive rates were 17.2%, 66.9%, 78.7%, 15.9%, and 0.14% for Clonorchis sinensis, Ascaris lumbricoides, Trichuris trichiura, hookworm, and Dicrocoelium dendriticum respectively. There was no significant difference between the infection rate of clonorchiasis, education level, and family income groups (p > 0.05). But there was significance difference between the infection rate of clonorchiasis and people living in different family sizes (p < 0.01). Thirty-four clonorchiasis patients treated with praziquantel 25 mg/kg/day for three days showed a cure rate in 30 days of 97.1%.

Infection status of Clonorchis sinensis in residents of Hamyang-gun, Gyeongsangnam-do, Korea

Oriental liver fluke (Clonorchis sinensis) infection was surveyed among residents of Hamyang-gun, Gyeongsangnam-do, Korea during the period of January 2001 to March 2002. Total 1,041 stool samples were collected from residents who visited Public Health Center and its branches in Hamyang-gun and examined using formalin-ether sedimentation method. The overall egg positive rate was 16%, male showing higher positive rate (21%) than female (10%). The age group of 30 to 50 years had the highest egg positive rate of C. sinensis from 20% to 22%. The positive examinees were treated with praziquantel and educated individually to prevent reinfection. Egg positive rate in this area was decreased when compared with results recorded in the past, however, still remained more than 10%. This study suggests that periodic examination, treatment as well as education of residents should be continued and systematized.

Epidemiology of clonorchiasis in Ninh Binh Province, Vietnam.

Clinical and stool examinations for clonorchiasis were carried out in an endemic area, Kim Son District, Ninh Binh Province, Vietnam. Stool examination with the Kato-Katz technic revealed that in 306 residents selected randomly, 42 people (13.7%) were infected with Clonorchis sinensis. The rate was biased towards men (23.4%) as opposed to women (1.5%) and increased with age. No children younger than 10 years old were infected, reflecting difference in a chance for acquisition of infection through a habit of eating raw fish. Few clinical abnormalities were found by blood and urine examinations of the patients. Treatment with praziquantel decreased the infection rate to 5.3% at 6 weeks later. Snails, Melanoides tuberculatus, collected from ponds around the settlements were infected with cercariae at a rate of 13.3%. Farmed fish (Hypophthalmichtys molitrix) in the ponds were infected with metacercariae at rates of 56.4% in small individuals and 100% in large ones. The life cycle of C. sinensis is exclusively completed in the ponds and the traditional habit of eating raw fish in summer was thought to be a major route of infection.

Antibodies in serum of patients with clonorchiasis before and after treatment.

Sera of 31 patients infected with Clonorchis sinensis were examined using fraction 1 antigen by ELISA during a one-year observation. The results of ELISA with Igs, IgG and IgA demonstrated high sensitivity (100%, 100% and 90%) and specificity (100%, 100% and 87%). Sera specific Igs and IgG were significantly decreased in the 3rd month after treatment with praziquantel (25mg/kg body weight in one dose), and IgA was significantly decreased in the 1st month (paired t-test, p < 0.05). No eggs were found in the stool after treatment. Detection of sera specific Igs, IgG and IgA by ELISA was combined with stool examination to evaluate the effect of praziquantel and the completeness of the cure.

Infecçäo por Clonorchis sinensis em imigrantes asiáticos no Brasil: tratamento com praziquantel / Clonorchis sinensis infections in Asian immigrants in Brazil: treatment with praziquantel

Estudaram-se quinze pacientes com infecçäo assintomática por Clonorchis sinensis, revelada através de exame parasitológico de fezes. Todos eram de origem asiática e procuraram o Laboratório Central do Instituto Adolfo Lutz para se submeterem a exames laboratoriais necessários a regularizaçäo de sua situaçäo, face a nova legislaçäo sobre imigrantes. Eram todos indivíduos adultos, seis pertencendo ao sexo feminino e nove ao masculino. Os quinze pacientes comn clonorquíase foram internados no Hospital das Clínicas da FMUSP e tratados com Praziquantel, na dosagem de 60 mg/Kg de peso corporal, dividida em duas tomadas. Foram realizados exames coprológicos quantitativos (método de Kato-Katz), antes do tratamento específico e no 15§, 30§ e 60§ dias após a terapêutica. Na última avaliaçäo (60§ dias após terapêutica), em nove pacientes (60,0%) näo se encontraram ovos do treamatódeo nas fezes e nos seis (40,0%), que continuavam eliminando ovos, notou-se reduçäo na quantidade eliminada (superior a 90% em cinco e a 30% no paciente restante). Os pacientes foram também submetidos a exames subsidiários, para avaliaçäo do estado geral e funçäo hepática, antes da administraçäo de Praziquantel e, posteriormente, no seguimento ambulatorial. A medicaçäo foi relativamente bem tolerada pelos pacientes, verificando-se a ocorrência de efeitos colaterais representados por náuseas e vômitos (dois casos), vertigens e tonturas (dois casos), epigastralgia (dois casos) e diarréia no 3§ dia após tratamento (um caso)